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刘若爽,董世魁,刘定震,吴永林,石建斌.2018.普氏原羚粪便样品中皮质醇激素保存时效研究.动物学杂志,53(1):131-137.
普氏原羚粪便样品中皮质醇激素保存时效研究
Storage Duration of Cortisol in Feces of Przewalski′s Gazelle (Procapra przewalskii)
投稿时间:2017-05-09  修订日期:2017-12-29
DOI:10.13859/j.cjz.201801017
中文关键词:  粪便样品  普氏原羚  皮质醇  低温保存
英文关键词:Fecal sample  Przewalski′s Gazelle (Procapra przewalskii)  Cortisol  Low temperature storage
基金项目:国家自然科学基金面上项目(No. 31572281),国家重大研发计划项目(No. 2016YFC0501906)
作者单位E-mail
刘若爽 北京师范大学环境学院 北京 100875 budthefox@escience.cn 
董世魁 北京师范大学环境学院 北京 100875 dongshikui@sina.com 
刘定震 北京师范大学生命科学学院 北京 100875 dzliu@bnu.edu.cn 
吴永林 青海湖国家级自然保护区管理局 西宁 810001 466952496@qq.com 
石建斌 北京师范大学环境学院 北京 100875 jbshi@bnu.edu.cn 
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中文摘要:
      由于采集粪便样品需对目标动物进行捕捉、限制或影响其行为,检测粪便样品中类固醇激素的方法近几十年来在野生动物保护、行为生态学、生理生态学等诸多领域得到广泛应用。普氏原羚(Procapra przewalskii)由于其种群和行为等方面的特殊性,较难进行直接的生理监测,因此采用间接手段监测其粪便类固醇激素就显得很有必要。为探讨不同保存方法对普氏原羚粪便中皮质醇的影响,我们将采集到的新鲜粪便样品充分混合后分为30份,各取10份分别保存于﹣20 ℃、4 ℃和20 ℃,在保存2、5、7、10、15、20、25、30 d和50 d时,使用酶联免疫吸附测定法(ELISA)检测样品中的皮质醇含量。单因素方差分析(One-way ANOVA)结果显示,保存于﹣20 ℃的粪便样品中皮质醇含量在保存50 d内没有显著变化,含量为(11.747 ± 2.951)ng/g(平均值 ± 标准差),(F = 1.966,n = 81,P > 0.05),而保存于4 ℃和20 ℃的样品则出现明显波动(4 ℃:F = 23.643,P < 0.05;20 ℃:F = 35.126,P < 0.05),含量分别为(15.951 ± 6.766)ng/g和(11.042 ± 6.094)ng/g。保存于4 ℃和20 ℃的样品中皮质醇含量在24 h内均有上升,在随后的几天逐渐下降。结果表明,﹣20 ℃冷冻可以简便有效地保存普氏原羚粪便样品中的皮质醇激素。同时,在野外暴露超过24 h的粪便样品会造成测定结果产生误差,在采集样品时要考虑到潜在的影响。
英文摘要:
      Because fecal samples can be collected without capturing and restraining animals, or disturbing animal behaviors, measurement of metabolites of steroid hormones by using feces has become very popular during the past decades, and it is currently being used in many areas of zoological research including wildlife conservation, behavioral ecology and physiological ecology. It is difficult to carry out direct physiological monitoring of Przewalski′s Gazelle (Procapra przewalskii) due to its particularity of population and behavior, so it is necessary to use the non-invasive sampling method to monitor the fecal steroids. We thus conducted this research to determine the effects of different storage temperatures on fecal cortisol in Przewalski′s gazelle. Fresh fecal samples were collected, thoroughly mixed and separated into 30 aliquots. Ten aliquots of mixed fecal samples were stored at ﹣20 ℃, 4 ℃ and 20 ℃, respectively. The cortisol concentrations in the samples were measured using ELISA on day 2, 5, 7, 10, 15, 20, 25, 30 and 50, respectively. The results of One-way ANOVA indicated no significant changes in the cortisol concentrations of the fecal samples stored at ﹣20 ℃ within 50 days (Mean ± SD = 11.747 ± 2.951 ng/g, F = 1.966, n = 81, P > 0.05). However, the cortisol concentrations of the fecal samples stored at 4 ℃ or 20 ℃ showed a significant fluctuation (4 ℃: Mean ± SD = 15.951 ± 6.766 ng/g, F = 23.643, P < 0.05; 20 ℃: Mean ± SD = 11.042 ± 6.094 ng/g, F = 35.126, P < 0.05). The cortisol concentration of the samples stored at 4 ℃ or 20 ℃ rose substantially after the first 24 h, and then declined after several days (Fig. 1). The results indicate that freezing at ﹣20 ℃ is a simple and reliable storage method for fecal cortisol in Prewalski′s gazelle. Meanwhile, because non-fresh fecal samples exposed in the field over 24 h may cause erroneous results, researchers should consider such potential effects when collecting fecal samples.
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